The growing use of telecommunication technologies by society has led the industry to develop infrastructure to support this growth. The telecommunications outdoor cabinets are an important element in the telecommunications network, allowing host and protect the equipment needed to run the network in adverse conditions. The enclosure must have a proper thermal performance to ensure a temperature below 55 � C, limit for the operation of equipment, and avoid an excessive consumption of electricity. This study aims to characterize the thermal performance of the cabinet using numerical studies with DesignBuilder/EnergyPlus. The development of the numerical model is followed by experimental studies using a real cabinet in operation, to ensure that the model is valid and allows to obtain reliable results. In the main results, it can be observed that the use of mechanical ventilation is effective in the extraction of heat from the interior of the cabinet, however, there is a
limit to which the increase in air flow rate does not result, significantly,…

► Higher derivative corrections to effective actions are very important and of great interest in string theory. The aim of this dissertation is to develop a…
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▼ Higher derivative corrections to effective actions are very important and of great interest in string theory. The aim of this dissertation is to develop a method to constrain the higher derivative corrections to O-plane actions using non-linear T-duality.
In this dissertation, we first illustrate this method with the simplest case without R-R field. We classify all possible two- and four-derivative couplings, which are compatible with diffeomorphism invariance and B-field gauge invariance, of bulk NS-NS sector fields with a single Op-plane. This is applicable to type IIA or IIB superstrings or to the bosonic string. We then consider this general action in various classes of backgrounds that admit a U(1) isometry and determine the constraints on the couplings from consistency with T-duality. We show that this consistency requires the two-derivative action to vanish, and the entire non-linear four-derivative action is fixed up to one overall constant which can be determined by comparison with a two-point scattering amplitude. The resulting action is consistent with all previously computed couplings. Then we use this method over actions involving any number of NS-NS fields and just one R-R field. We first list all possible couplings up to four derivatives, then we reproduce the T-duality procedure for two-derivative case and show that the action vanishes in this case, which is also consistent with results in literature.
Advisors/Committee Members: Becker, Katrin (advisor), Becker, Melanie (committee member), Kamon, Teruki (committee member), Fulling, Stephen A (committee member).

McLagan, T. L. (2017). Re[growth] :
a development of social capital through food network
accessibility. (Masters Thesis). University of Pretoria. Retrieved from http://hdl.handle.net/2263/60188

Chicago Manual of Style (16th Edition):

McLagan, Toni Lisa. “Re[growth] :
a development of social capital through food network
accessibility.” 2017. Masters Thesis, University of Pretoria. Accessed May 25, 2019.
http://hdl.handle.net/2263/60188.

MLA Handbook (7th Edition):

McLagan, Toni Lisa. “Re[growth] :
a development of social capital through food network
accessibility.” 2017. Web. 25 May 2019.

Vancouver:

McLagan TL. Re[growth] :
a development of social capital through food network
accessibility. [Internet] [Masters thesis]. University of Pretoria; 2017. [cited 2019 May 25].
Available from: http://hdl.handle.net/2263/60188.

Council of Science Editors:

McLagan TL. Re[growth] :
a development of social capital through food network
accessibility. [Masters Thesis]. University of Pretoria; 2017. Available from: http://hdl.handle.net/2263/60188

► Many proteins of eukaryotic cells are known to be O-glycosylated. Glycoproteins with heavily O-glycosylated mucin domains provide important biological functions in a cell: i.e., protection…
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▼ Many proteins of eukaryotic cells are known to be
O-glycosylated. Glycoproteins with heavily O-glycosylated mucin
domains provide important biological functions in a cell: i.e.,
protection from pathogens, cell-to-cell adhesion and intracellular
protein trafficking. Mucin-type O-glycosylation occurs in the Golgi
complex and begins with the transfer of GalNAc from UDP-GalNAc onto
Ser/Thr residues of polypeptides. This step is catalyzed by a large
family (20) called N-a-acetylgalactosaminyl transferases
(ppGalNAc-T’s) and forms the GalNAc-a-O-Ser/Thr product. Subsequent
elongation is performed by specific glycosyltransferases, producing
a variety of glycans. Family members have been classified into
peptide- and glycopeptide-preferring subfamilies, although both
subfamilies possess variable activities against glycopeptide
substrates. Structurally, 19 isoforms contain a C-terminal
catalytic domain linked via a flexible linker to an N-terminal
ricin-like lectin domain. The (glyco)peptide substrate
specificities of the ppGalNAc-T transferases and the roles of the
catalytic and lectin domains in glycopeptide glycosylation still
remain largely unknown. Based on the systematic random peptide
approach created by the Gerken Lab, I have determined the
glycopeptide substrate specificities of several ppGalNAc-T
isoforms. A series of (glyco)peptides were created in order to
specifically probe the functions of the catalytic and lectin
domains in terms of neighboring (1-5 residues) and remote prior
glycosylation (6-17 residues) from an acceptor site, respectfully.
Using several glycopeptide-preferring isoforms, glycosylation was
observed from -4, -3, -1 and +1 relative to a neighboring
GalNAc-O-Thr, which I attributed to specific GalNAc-O-Thr binding
at the catalytic domain. The other series of glycopeptides
contained a GalNAc-O-Thr near the C- or N- terminus of the
substrate to address the directionality preferences of the lectin
domain. Results with several peptide- and glycopeptide-preferring
isoforms revealed preferences that varied among transferase
isoform, where some preferred a C-terminally placed GalNAc-O-Thr,
or a N-terminally placed GalNAc-O-Thr and others equally preferred
the C-/N- terminally placed GalNAc-O-Thr. These directionality
preferences are due to the GalNAc-O-Thr interactions at the lectin
domain. Results of these studies revealed for the first time the
site-specific glycopeptide glycosylation preferences by some
ppGalNAc-T’s and has demonstrated that both domains of the
ppGalNAc-T’s have specialized and unique functions that work in
concert to control and order mucin-type
O-glycosylation.
Advisors/Committee Members: Gerken, Thomas (Advisor), Lee, Irene (Committee Chair).

► CD46 is a ubiquitously expressed transmembrane protein in humans with a role in immune homeostasis. Originally identified as a complement regulator, CD46 has since been…
(more)

▼ CD46 is a ubiquitously expressed transmembrane protein in humans with a role in immune homeostasis. Originally identified as a complement regulator, CD46 has since been regarded a receptor for several pathogens and most recently, described as a T cell costimulatory molecule. Its coligation with CD3 and consequent cleavage from the T cell surface serves as a costimulatory stimulus for T cell activation. In addition, in the presence of IL-2, CD46 induces Tr1 cell differentiation which is characterised by low IFN-γ and high IL-10 secretion. CD3/CD46-induced Tr1 differentiation is defective in patients with MS, rheumatoid arthritis and asthma, highlighting the need to investigate the mechanisms involved in the regulation of the CD46 pathway. CD46 is a highly glycosylated protein with three N-glycosylation sites in the short consensus repeats and multiple O-glycosylation sites in the STP region. Previous data from the lab have shown that CD3 activation causes a change in CD46 glycosylation. Herein, I convey that this change is more pronounced in memory than naive CD4+ T cells and is mainly due to changes in CD46 O-glycosylation. Furthermore, these changes are required for the T cell responses triggered by CD46 costimulation including T cell activation and Tr1 differentiation. Interestingly, CD46 is recruited to the immunological synapse formed between a T cell and an antigen presenting cell and I illustrate that the STP region is needed for this also. These data suggest that the glycosylation status of CD46 regulates its function. In MS, vitamin D deficiency is considered to be a significant risk factor and many patients take vitamin D supplement to help manage their condition. Herein, I report that treatment of healthy and MS CD4+ T cells with vitamin D does not prevent T cell activation but it decreases adhesion molecule expression. Moreover, vitamin D supplementation in MS enhances CD46 cleavage. Therefore, vitamin D also plays a role in the regulation of the CD46 pathway and it would be interesting to investigate whether vitamin D affects CD46 glycosylation. During my MSc, I showed that a recombinant protein derived from adenovirus serotype 35 (which naturally binds CD46) known as Ad35K++ controls the CD46 pathway in CD4+ T cells. Lymphoma cells treated with Ad35K++ in combination with monoclonal antibody therapy rituximab have demonstrated increased sensitivity to rituximab and prove that virus-derived recombinant proteins that target CD46 have therapeutic potential. Considering the key role of CD46 as a T cell costimulatory molecule, I have investigated the effects of Ad35K++ on the CD46 pathway following its use in vivo and confirm CD46 is still cleaved from the cell surface and the cells still become activated. Overall these results provide insight into the mechanisms involved in the regulation of the CD46 pathway and highlight how it can be manipulated for therapeutic use.

Niobium based refractory intermetallic alloys are considered as having great potentials for high temperature applications based on a good balance of high temperature strength and low-temperature damage tolerance. Moreover this family of alloys also exhibits a high melting point and a low density; hence they are good candidates for low pressure turbine blade applications over the temperature range of 800–1000 °C. The aim of this study is to investigate the chemical composition changes, especially the O-Ti2AlNb phase precipitation and the silicon addition effects on the microstructures and high temperature mechanical properties. This study starts with a first prospective on the chemical compositions of alloys showing the highest potential for applications wanted. The studied alloys show a good balance between the ductility at room temperature induced by titanium which stopped the fragile intermetallic d-Nb3Al precipitation; and a high niobium content to maintain good high temperature mechanical properties. Two alloys are doped with 1 %at Si to improve high temperature strength and keep an acceptable ductility at room temperature. The result indicates on the more promising alloy chemical composition. We also investigate the influence of aluminum and silicon…

► The synthesis of pure tris(6-hydroxymethyl-2-pyridylmethyl)amine (H₃L₁₁) is reported for the first time. New complexes of H₃L₁₁ with copper(II), manganese(II) and iron(III) have been characterised by…
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▼ The synthesis of pure tris(6-hydroxymethyl-2-pyridylmethyl)amine (H₃L₁₁) is reported
for the first time. New complexes of H₃L₁₁ with copper(II), manganese(II) and iron(III) have been characterised by X-ray crystallography. Linear [Fe₃(L₁₁)₂](ClO₄)₃ reveals the tightest Fe-O-Fe angle (87.6°) and shortest Fe...Fe distance (2.834 Å) presently found for a weakly antiferromagnetically-coupled high spin alkoxide-bridged polyiron(III) system.
H₃L₁₁ provides a route to various hydrophobic peralkylated TPA ligand derivatives for creating a hydrophobic pocket for the assembly of iron catalysts for the novel 1-hydroxylation of n-alkanes. New 6-py substituted TPA ligands containing methyl (L₁₅) and n-octyl (L₁₆) ether linkages were synthesised via alkylation. Two further novel 6-py substituted ligands were synthesized incorporating n-hexyl substituents on one (L₂₁) and two (L₂₂) of the py moieties. Here a urea spacer group was used to promote hydrogen–bond assisted heterolytic O-O cleavage (generation of the potent FeV=O oxidant) within the hydroxoperoxoiron(III) precursor. High spin [FeII(L)(CH₃CN)[subscript(x)]](CF₃SO₃)₂ complexes (x = 0–2, L = L₁₅,₁₆,₂₁,₂₂) were characterised in solution by ¹H NMR. The structure of [Fe(L₂₂)](CF₃SO₃)₂ reveals a distorted iron(II) centre bound to four N atoms and two urea carbonyls.
Iron(II) complexes of H₃L₁₁, L₁₅,₁₆,₂₁,₂₂ and tris(6-Br)-TPA (L₂₄), were investigated for catalysis of the oxygenation of cyclohexane by H₂O₂. Reaction of the iron(II) complexes with H₂O₂ and [superscript(t)]BuOOH was followed by time-resolved EPR and UV-VIS spectrophotometry. A correlation between the observed catalytic activity and the nature of the FeIII(L)-OOR intermediates generated is apparent.
A convenient ‘one-pot’ synthesis of benzene-1,3,5-triamido-tris(l-histidine methyl ester) is reported along with attempts at preparing N,N’-bis(pyridylmethyl)-1,3- diaminopropane-2-carboxylic acid (L₂₅), a new water soluble pyridine-amine ligand. The final demetallation step resulted in ligand hydrolysis to the novel amino acid; 1,3-diaminopropane- 2-carboxylic acid which was characterised as its HCl salt by X-ray crystallography.
Advisors/Committee Members: Richens, David. T (advisor).

► The premise upon which this thesis is founded is that the book of Qohelet is fundamentally ambiguous. Ambiguity is attached to all its major themes,…
(more)

▼ The premise upon which this thesis is founded is that the book of Qohelet is fundamentally ambiguous. Ambiguity is attached to all its major themes, and can be discerned in its language, syntax and structure. This has not been given due attention in previous works on Qohelet.
The introduction considers the concepts of 'ambiguity' and 'meaning': it is crucial for the reader to understand what is meant in this thesis by these terms. 'Ambiguity' is
understood as those aspects of the text whose indeterminacy requires the reader to fill in 'meaning' in order for a coherent reading to be produced: thus the reader's role is crucial, but is nonetheless restricted by the determinate schemata in the text.
Part 1 explores the determinate schemata in Qohelet in an attempt to provide objective
criteria against which the ambiguities may be set. Detailed attention is paid to the text in order to discern trends and patterns in the book. These are employed in an attempt to discover how the book as a whole and the sections within it are structured. Part 1 ends by asserting that it is ultimately futile to seek an overall structure or pattern to the book: this is an aspect of its ambiguity.
Part 2 systematically examines linguistic and syntactical ambiguities in Qohelet, exploring the possibilities for interpretation according to the ways in which the reader fills in the gaps left by these ambiguities.
The conclusion argues that the ambiguity of Qohelet is the primary reason for the hugely diverse interpretations of the book throughout its history, and for the many varied proposals for its structure. In this way it is a realistic reflection of an ambiguous world and the relationship between the people of this world and the God who made the world with all its ambiguities.

Ingram, D. N. (1996). The ambiguity of Qohelet : a study of the ambiguous nature of the language, syntax and structure of the Masoretic text of Qohelet. (Doctoral Dissertation). University of Stirling. Retrieved from http://hdl.handle.net/1893/2589

Chicago Manual of Style (16th Edition):

Ingram, Douglas Nairn. “The ambiguity of Qohelet : a study of the ambiguous nature of the language, syntax and structure of the Masoretic text of Qohelet.” 1996. Doctoral Dissertation, University of Stirling. Accessed May 25, 2019.
http://hdl.handle.net/1893/2589.

MLA Handbook (7th Edition):

Ingram, Douglas Nairn. “The ambiguity of Qohelet : a study of the ambiguous nature of the language, syntax and structure of the Masoretic text of Qohelet.” 1996. Web. 25 May 2019.

Vancouver:

Ingram DN. The ambiguity of Qohelet : a study of the ambiguous nature of the language, syntax and structure of the Masoretic text of Qohelet. [Internet] [Doctoral dissertation]. University of Stirling; 1996. [cited 2019 May 25].
Available from: http://hdl.handle.net/1893/2589.

Council of Science Editors:

Ingram DN. The ambiguity of Qohelet : a study of the ambiguous nature of the language, syntax and structure of the Masoretic text of Qohelet. [Doctoral Dissertation]. University of Stirling; 1996. Available from: http://hdl.handle.net/1893/2589

With the latest reforms, among which the Hospital, Patient, Healthcare and Territory (HPST) Law, the French Hospital has changed not only the form of its organisation through the implementation of a polar organisation, but also its environmental concern. This polar organisation aims to strive for performance and efficiency via an managerial and medical approach. Thus, some management tools have been set up aimed in particular at giving feedback on such performance. In this context, how can professionals from such distinct fields as management and healthcare work? Management implies the optimization of resources when the public sector above all upholds values of social usefulness and patient care. Healthcare actors feel very much involved in this mission of social usefulness. In this work we propose to identify how cultural gaps within hospital poles contribute to a loss of meaning and acknowledgment among healthcare agents. Those cultural gaps will be examined more particularly through management tool appropriation. This study relies on interviews conducted in various healthcare facilities (PCO and psychiatric hospitals of various capacities such as CHU and CH). It reveals the need for innovative managerial practices, especially…

► The objective of this paper is to analyze the different portfolios formed by Markowitz's (1952) mean-variance model and a naive strategy (1/N) composed of ETFs…
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▼ The objective of this paper is to analyze the different portfolios formed by Markowitz's (1952) mean-variance model and a naive strategy (1/N) composed of ETFs that can surpass the Bovespa Index. We used 13 ETFs listed on the S??o Paulo Stock Exchange from 01/01/2012 to 04/11/2016 to perform four portfolio optimization and weighting strategies: 1) Model with short sale and leverage restrictions; 2) Portfolios with a maximum position of 30% in each asset; 3) Wallets with a leveraged position, where it is allowed to be sold up to -30% and bought up to 130% in each asset (strategy 130/30); 4) Portfolios where each asset may be short-listed up to a limit of -5%, provided that the maximum percentage sold in the portfolio does not exceed -45%. For each of these strategies, balances were determined in temporal windows of 1, 3, 6 and 12 months. In all analyzes, there was no significant difference in terms of return, although naive portfolios (1/N) had marginally higher numbers. Optimized portfolios composed of ETFs in the different strategies presented a significantly lower risk than the naive portfolio of ETFs and the market index. There was no predominance in terms of risk and return in the comparison between naive portfolios composed of ETFs and naive portfolios composed of randomly chosen Stock Investment Funds (FIAs). ETFs feature ease of trading, transparency and economy, and can become an alternative in the composition of the investor's asset portfolio.
Advisors/Committee Members: Contani, Eduardo Augusto do Ros??rio (advisor), http://lattes.cnpq.br/9581308995467871 (advisor).

► The objective of this study is to analyze the factors that determine the spread of the public issues of debentures indexed to the Broad Consumer…
(more)

▼ The objective of this study is to analyze the factors that determine the spread of the public issues of debentures indexed to the Broad Consumer Price Index (IPCA) in Brazil. Emissions indexed to the IPCA were choose because they are instruments usually used to capture resources of longer maturity by the issuing companies. The database had 245 series of issues occurred between January 2010 and December 2015. Regressions were estimated by ordinary least squares and weighted least squares methods, and the results presented by the last method were more robust. The rating-spread ratio was confirmed in all regressions and the results indicate that this variable explains, by itself, 58% of the spread variation. Other the rating, the results indicate that the main factors that determine the spread of the issues are: collateral, issuer experience, maturity, amount, prestige of the coordinating bank, tax benefits and economic scenario. Due to the results achieved, other issues related to market efficiency were approached, such as agency conflicts, information asymmetry and adverse selection. The results show that the collaterized issues remunerated the investors with a higher spread than the unsecured ones, and this premium ranged from 35 to 38 basis points. The results were interpreted in the context of agency theory and resemble those found by John, Lynch and Puri (2003) for the US corporate bonds market. Finally, the favorable economic scenario, as measured by the Emerging Market Bond Index - Brazil (EMBI + BR), showed negativelycorrelated with the spread, and these results were interpreted as effects of the information asymmetry and adverse selection present in the local market for debt issuance.
Advisors/Committee Members: Yoshinaga, Claudia Emiko (advisor), http://lattes.cnpq.br/9991689534310081 (advisor).

The present report will describe all the activities that I have performed during my 6 months internship as a master student of clinical analysis at the Faculty of Pharmacy of the University of Lisbon. I worked as an intern at Nova Era-Luz laboratory and I did several procedures in three specific areas: Biochemistry, Hematology and Immunology. At the beginning, in the biochemistry section, I started by doing serum (Olympus AU400 e MiniCap) and urine (Aution Max AX4280 – chemical parameters) analyses. Hematology department is the one that has a major number of manual techniques and automatic devices which include Sysmex XT2000i (CBC); VesMatic 30/30 Plus (sedimentation rate); Adams A1C HA8160 (glycated hemoglobin assay); Hb Gold (hemoglobin electrophoresis) and Sysmex Ca500 (blood coagulation assay). Lately, in the immunology section I took contact with the two principle devices that are used for this type of analysis: Advia Centaur and VIDAS. So, this report is divided
in the three sections mentioned above and in which one of them I present a theoretical foundation of the technique, principles and an explanation of the obtained results either by device or manual technique.

GILZ (Glucocorticoid-Induced Leucine Zipper) and TSC-22 (Transforming growth factor-beta Stimulated Clone-22) belong to the TSC-22D (TSC-22 Domain) family of proteins. GILZ has been previously shown to be induced upon interleukin-2 (IL-2) deprivation in the T-cell line CTLL-2, allowing cells to delay apoptosis. The aim of our study was to elucidate the respective roles of GILZ and TSC-22 during IL-2 deprivation-induced T-lymphocytes apoptosis.Our results demonstrated that TSC-22 increased CTLL-2 cells apoptosis induced upon IL-2 deprivation. We highlighted in TSC-22 expressing cells both an increase in caspases activation and BIM expression up-regulation. We also demonstrated that GILZ expression, an anti-apoptotic protein, known to be induced after IL-2 withdrawal, was down-regulated in the presence of TSC-22. Moreover, we showed that gilz mRNA expression was also significantly repressed, but gilz mRNA half-life was not modified.Altogether, these results suggest that, in T-cells, TSC-22 could behave as a repressor of GILZ expression, accelerating IL-2 deprivation-induced apoptosis.

Listeria monocytogenes is a foodborne pathogen responsible for human listeriosis which invade and replicate in both phagocytic and non-phagocytic cells. Listeria intracellular life cycle involves interference with host cell components. My PhD project aims to characterize host cell proteome modifications in cells infected by Listeria. I focused my research on host proteases activation and the degradation of host cell proteins in response to infection. A proteomic analysis performed on cells treated with purified Listeriolysin O (LLO), a pore forming toxin secreted by Listeria, identified more than 90 proteins degraded in response to the toxin. We validated the degradation of these proteins using both in…

►O-linked β-N-acetylglucosamine (O-GlcNAc) is a ubiquitous post-translational protein modification found on serine and threonine amino acid residues of intracellular proteins. This inducible and dynamic PTM…
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▼O-linked β-N-acetylglucosamine (O-GlcNAc) is a ubiquitous post-translational
protein modification found on serine and threonine amino acid residues of intracellular proteins. This inducible and dynamic PTM is mediated by two cycling enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) in response to environmental stimuli. The nucleotide-sugar donor UDP-GlcNAc is the end product of the hexosamine biosynthetic pathway (HBP) and is responsive to glucose levels entering the cell. Provided its proposed role as a cellular nutrient sensor, O-GlcNAc has been implicated in contributing to the progression of type II diabetes. However, the molecular role for this PTM in the glucose-responsive, insulin secreting pancreatic β cell remains unclear. In this dissertation, I set out to study the role of O-GlcNAc in regulating molecular events in the β cell, specifically at the levels of insulin secretion and transcription. Using directed pharmacological approaches in the mouse insulinoma-6 (Min6) cell line, I demonstrate that elevating nuclear O-GlcNAc preserves glucose stimulated insulin secretion during chronic hyperglycemia. This observed secretory effect directly correlates with O-GlcNAc-induced elevation in perinuclear insulin under basal and prolonged hyperglycemic conditions. The molecular mechanism for these observed changes appears to be, at least in part, due to elevated O-GlcNAc-dependent increases in Ins1 and Ins2 mRNA levels via elevations in histone H3 transcriptional activation marks. Further, whole transcriptome shotgun sequencing reveals that hyperglycemia altered gene transcription is restricted to a subset of genes and that the majority of genes regulated by inhibiting OGA levels are similarly regulated by a shift from euglycemic to hyperglycemic conditions. Thus, my work demonstrates a role for O-GlcNAc as a glucose sensor and modulator of gene transcription in pancreatic β cells.
Advisors/Committee Members: Lance Wells.

Durning SP. O-GlcNAc is a sensor in the pancreatic beta cell. [Internet] [Doctoral dissertation]. University of Georgia; 2013. [cited 2019 May 25].
Available from: http://purl.galileo.usg.edu/uga_etd/durning_sean_p_201312_phd.

Council of Science Editors:

Durning SP. O-GlcNAc is a sensor in the pancreatic beta cell. [Doctoral Dissertation]. University of Georgia; 2013. Available from: http://purl.galileo.usg.edu/uga_etd/durning_sean_p_201312_phd

►O-linked mannose has been implicated in cell-cell and cell-matrix adhesion. Defects in glycosyltransferases that add or extend O-linked mannose on glycoproteins generate neuron migration defects…
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▼O-linked mannose has been implicated in cell-cell and cell-matrix adhesion. Defects in glycosyltransferases that add or extend O-linked mannose on glycoproteins generate neuron migration defects in congenital muscular dystrophies (CMD). The only well characterized O-mannose modified mammalian protein has been the peripheral membrane protein alpha-dystroglycan (±-DG). We propose that there must be other O-mannosylated proteins that modulate the CMD phenotype since O-mannosylation comprises a third of all O-linked glycans in the brain, and a brain ±-DG knockout does not recapitulate all neuronal aspects of CMD. This research proposal aims to develop a chemoenzymatic labeling method to identify those proteins. Elucidating the functional role of O-mannose would contribute towards existing knowledge of how extracellular glycosylation participates in a broad range of human disorders and tissue functions while also paving the way for glycopeptide and enzyme based therapeutics. KEYWORDS: O-Mannose, Neuron Migration, Chemoenzymatic Labeling USING CHEMOENZYMATIC LABELING TO PROBE O-MANNOSE GLYCOSYLATION By ANURADHA R. KOPPIKAR B.S., Georgia Institute of Technology, 2005 A Thesis Submitted to the Graduate Faculty of The University of Georgia in Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE ATHENS, GA 2008
Advisors/Committee Members: Lance Wells.

▼ Insulin resistance defines the metabolic syndrome and precedes type 2 diabetes, thus is considered as the hallmark for type 2 diabetes. Prolonged hyperglycemia and hyperinsulinemia are both required for the development of classical insulin resistance. In recent years, accumulative evidence has established that elevated O-GlcNAc levels via either genetic or pharmacological methods lead to insulin resistance in both cultured adipocytes and animal models. Adipocytes, besides being a major site for energy storage, are endocrine in nature and secrete a variety of proteins (adipocytokines) that can modulate insulin sensitivity, hypertension, food intake, and general energy homeostasis. The link between O-GlcNAc levels, insulin resistance, and adipocytokine secretion is further explored in this dissertation.
First, by using immortalized and primary rodent adipocytes, the secreted proteome of
differentiated adipocytes under insulin responsive and two insulin resistant conditions is
explicitly elucidated via shotgun proteomics leading to the identification of 97 and 203 secreted proteins, respectively. More than 80 N-linked glycosylation sites on adipocytokines released by these adipocytes were also mapped. Moreover, we took a step further to address the quantitative secretome of human primary adipocytes with the same technique. Of the 190 secreted proteins identified, we report 20 up-regulated and 6 down-regulated proteins detected in both insulin resistant conditions. Moreover, we applied glycomic techniques to examine (1) the N-linked sites from the pool of secreted proteins and (2) the characterization and relative abundance of complex N- and O-glycans released from adipocytes exposed to different conditions. We identified 91 Nglycosylation sites on the secreted proteins derived from 51 proteins, as well as 155 and 29 released N- and O-glycans respectively. There were moderate alterations in the observed N- and O-linked glycans under the different conditions considered.
In short, our studies have provided a list of adipocytokines whose secreted levels vary
under insulin resistant conditions. These proteins bear the potential to be prognostic/diagnostic biomarkers for the detection of metabolic syndrome and type 2 diabetes. Also, comparing the results complied from both classical and non-canonical insulin resistance provides further insights for the role of the O-GlcNAc modification on intracellular proteins in the disease progression of type 2 diabetes.
Advisors/Committee Members: Lance Wells.

►O-GlcNAc transferase (OGT) is responsible for the addition of the β-N-acetylglucosamine post-translational modification to serine/threonine residues of hundreds of nuclear and cytoplasmic proteins. In a…
(more)

▼O-GlcNAc transferase (OGT) is responsible for the addition of the β-N-acetylglucosamine post-translational modification to serine/threonine residues of hundreds of nuclear and cytoplasmic proteins. In a focused X chromosome exome next generation sequencing of 30 probands with X-linked Intellectual disability (ID), a novel missense mutation in the OGT gene (Xq13.1) has been identified in a family with three affected males. The mutation occurs in the tetratricopeptide(TPR) region [762G>T (p.L254F)]of the transferase. The clinical phenotypes of these patients include hypospadia, clinodactyly, short stature, microcephaly, and- ID. To study the physiological role of this mutation, lymphoblastoid cell lines from two affected males, one mother and three unaffected related males were isolated. Steady-state OGT protein levels are decreased in the patient samples compared to the carrier and normal control in agreement with molecular modeling that predicts the mutation to be destabilizing. This was further validated by half-life studies that demonstrate a faster turnover of the L254F-OGT. We have generated a recombinant L254F-OGT that has allowed us to perform activity studies and L254F-OGT is active in vivo against protein substrates and in vitro against a synthetic peptide. Surprisingly, steady-state global O-GlcNAc levels remain grossly unaffected in XLID. The same samples, however, show a decrease in steady-state O-GlcNAcase (OGA, the enzyme that removes O-GlcNAc from proteins) levels. These findings imply a compensation mechanism exists, although imperfect, given the phenotype of the patients, for maintaining global O-GlcNAc levels. L254F-OGT patients also show a decrease in OGA steady state mRNA levels and luciferase reporter expression. OGT has been previously shown to exist in a co-repressor complex to down regulate gene expression. We have observed that there is enrichment of OGT at the proximal promoter region of OGA leading us to hypothesize that OGT regulates OGA transcription in the patient lymphoblastoids. In parallel, global transcriptome analysis by performing RNA deep sequencing has revealed that there are disease specific changes in gene expression. Currently we are determining the mechanism of regulation of OGA gene repression by OGT as well as validating targets obtained by RNA deep sequencing analysis. Finally, we will generate induced pluripotent stem (iPS) cells from the fibroblast of affected patients and carriers in order to explore the imperfect compensatory mechanism in derived neural lineages due to the specific phenotypes observed. For the first time, we have identified and partially characterized a missense mutation in OGT that causes a disease, XLID.
Advisors/Committee Members: Lance Wells.

Vaidyanathan, K. (2014). Identification and characterization of a missense mutation in O-GlcNAc transferase that segregates with disease in a family with X-linked intellectual disability. (Doctoral Dissertation). University of Georgia. Retrieved from http://purl.galileo.usg.edu/uga_etd/vaidyanathan_krithika_201405_phd

Chicago Manual of Style (16th Edition):

Vaidyanathan, Krithika. “Identification and characterization of a missense mutation in O-GlcNAc transferase that segregates with disease in a family with X-linked intellectual disability.” 2014. Doctoral Dissertation, University of Georgia. Accessed May 25, 2019.
http://purl.galileo.usg.edu/uga_etd/vaidyanathan_krithika_201405_phd.

MLA Handbook (7th Edition):

Vaidyanathan, Krithika. “Identification and characterization of a missense mutation in O-GlcNAc transferase that segregates with disease in a family with X-linked intellectual disability.” 2014. Web. 25 May 2019.

Vancouver:

Vaidyanathan K. Identification and characterization of a missense mutation in O-GlcNAc transferase that segregates with disease in a family with X-linked intellectual disability. [Internet] [Doctoral dissertation]. University of Georgia; 2014. [cited 2019 May 25].
Available from: http://purl.galileo.usg.edu/uga_etd/vaidyanathan_krithika_201405_phd.

Council of Science Editors:

Vaidyanathan K. Identification and characterization of a missense mutation in O-GlcNAc transferase that segregates with disease in a family with X-linked intellectual disability. [Doctoral Dissertation]. University of Georgia; 2014. Available from: http://purl.galileo.usg.edu/uga_etd/vaidyanathan_krithika_201405_phd

University of Georgia

29.
Brimble, Sandra Nicole.
Investigating the role of O-GlcNAc in the regulation of human Oct4.

►O-linked β-N-acetylglucosamine (O-GlcNAc) is a single sugar modification found on many different classes of nuclear and cytoplasmic proteins. Addition of this modification, by the enzyme…
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▼O-linked β-N-acetylglucosamine (O-GlcNAc) is a single sugar modification found on many different classes of nuclear and cytoplasmic proteins. Addition of this modification, by the enzyme O-linked N-acetylglucosamine transferase (OGT), is dynamic and inducible. There is mounting evidence that O-GlcNAc plays a role in regulation of development but the mechanism is not clearly understood. One major class of proteins modified by O-GlcNAc is transcription factors. O-GlcNAc regulates transcription factor properties through a variety of different mechanisms including localization, stability and transcriptional activation. Maintenance of embryonic stem (ES) cell pluripotency requires tight regulation of several key transcription factors, many of which are modified by O-GlcNAc. Pou5f1 (Oct4) is one of the transcription factors required for pluripotency of ES cells and more recently, the generation of induced pluripotent stem (iPS) cells. The action of Oct4 is modulated by the addition of several post-translational modifications, including O-GlcNAc. Previous studies in mouse found a single site of O-GlcNAc addition responsible for transcriptional regulation. This study was designed to determine if this mechanism is conserved in human. We mapped 10 novel sites of O-GlcNAc attachment on human Oct4, and confirmed a role for OGT in transcriptional activation of Oct4 at a site distinct from that found in mouse that allows distinction between different Oct4 promoters. Additionally, we uncovered a potential new role for OGT that does not include its catalytic function. These results confirm that human Oct4 activity is being regulated by OGT by a mechanism that is independent of O-GlcNAc and distinct from mouse Oct4.
Advisors/Committee Members: Lance Wells.

Brimble, S. N. (2014). Investigating the role of O-GlcNAc in the regulation of human Oct4. (Doctoral Dissertation). University of Georgia. Retrieved from http://purl.galileo.usg.edu/uga_etd/brimble_sandra_n_201408_phd

Chicago Manual of Style (16th Edition):

Brimble, Sandra Nicole. “Investigating the role of O-GlcNAc in the regulation of human Oct4.” 2014. Doctoral Dissertation, University of Georgia. Accessed May 25, 2019.
http://purl.galileo.usg.edu/uga_etd/brimble_sandra_n_201408_phd.

MLA Handbook (7th Edition):

Brimble, Sandra Nicole. “Investigating the role of O-GlcNAc in the regulation of human Oct4.” 2014. Web. 25 May 2019.

Vancouver:

Brimble SN. Investigating the role of O-GlcNAc in the regulation of human Oct4. [Internet] [Doctoral dissertation]. University of Georgia; 2014. [cited 2019 May 25].
Available from: http://purl.galileo.usg.edu/uga_etd/brimble_sandra_n_201408_phd.

Council of Science Editors:

Brimble SN. Investigating the role of O-GlcNAc in the regulation of human Oct4. [Doctoral Dissertation]. University of Georgia; 2014. Available from: http://purl.galileo.usg.edu/uga_etd/brimble_sandra_n_201408_phd

▼ Latina/o health discourses stem from historical and social notions of biological, cultural, and racial inferiority. Popular U.S. newspapers pay scant attention to Latina/o health concerns and often inaccurately portray Latinas/os as undeserving foreigners that continue to drain social services such as health care. A content analysis of 291 New York Times, Los Angeles Times, Chicago Tribune, and Houston Chronicle newspaper articles (2006-2010) reveals that Latina/o health discourses are grounded in a racialized medical narrative that justifies and sustains white racial oppression. Systemic racism and the white racial frame are utilized as theoretical frameworks to better understand how mainstream newspapers construct the medical racialization of Latinas/os and contribute to health disparities, unequal access to health services, and inadequate health care. The findings reveal that Latina/o health issues concerning high costs, population increase, and political marginality, influence anti-Latina/o legislation, sustain prevailing racism, and create exclusionary health practices. Fundamentally the anti-Latina/o sentiment presented in the newspapers and disseminated throughout society equates to the denial of resources, the denial of health care, and thus the denial of life. Challenging racist Latina/o perceptions is an important area of social science and anti-racism research. Ultimately, without a healthy Latina/o workforce, the economy could not sustain itself and society would be susceptible to economic, social, and political collapse.
Advisors/Committee Members: Feagin, Joe (advisor).